Ferm Utilities
What do we cool our fermenters with and what is the temperature set point?
RO water in a closed loop that feeds the chillers at a SP of 35*F
What temperature and time are needed to sterilize the media and dextrose through the holding loops? What calculation is used to determine these values? And what is our target value for this calculation to determine if our system is sterile?
265*F for 6min
F0 calculation = 40
-80*C and forever
What do we add to our fermentation vessels to maintain level of the tank and to prevent it from going out the wall?
Bonus 100 points: what is the name of this chemical (not just the acronym)
antifoam
PPG-2000 - polypropylene glycol 2000
Define what DO is and why it is important.
DO - dissolved oxygen, measurement of how much oxygen is in a liquid
How many sections of cooling jackets are on the fermenters? And what is the purpose of cooling the tanks? When is cooling most needed?
8 sections
Purpose: cool the vessel to inoc with out killing org, and org creates exothermic reaction and constantly needs to be cooled.
Cooliing is most needed within the first 5 horus of fermentation
Name all of the media ingredients into the media sterilizer. Why cannot some of them mix?
RO water, Group 1: mag sulfate, mag soln, trace metals, Group 2: DKP, phos acid, citric acid
what is the purpose of the seed fermenter? What is the target DO and why is that important?
replicate and produce biomass
target DO is 20%+ aerobic fermentation for optimal growth
What is the purpose of the production vessel? What is the target DO and why is that important?
produce BDO
target DO is <2% and goes into micro-aerobic state to stress the organism out and push it to BDO production.
What is the purpose of a scrubber? How many different types of scrubbers have and how do they work?
purpose: contain organism from going out into the atmosphere, also scrubs VOCs
- cyclonic: cyclonic action with water to capture the organism and send it down the drain
wet: has packing, captures org through packing and drops it down into drain of scrubber
Why do we feed our fermenters with RO water? Why not process water?
RO removes all of the ions in the water so there are no added ions. We add a very specific recipe in our fermenters and cannot have a variable addition of ions feeding the fermenters.
Why is ammonia added after the sterilization holding loop? And how is it sterile?
Do not want ammonia to vaporize as it is a very volatile compound. Also cannot consume the nitrogen in a vapor form.
After cryobullet, where does our organism transition to? How long is it in this vessel? And what parameter tells us to move to the next step?
BONUS for 100 points: what value is the parameter that tells us to move it forward to the next step?
shake flask, 17hrs, OD - optical density
bonus - 6 OD
Identify all fermentation systems that are supplied by the fermentation CIP system. There need to be 6 responses for points.
1. SFs/PFs
2. transfer header
3. Inoculation header
4. broth surge tanks
5. broth kill sterilizer
6. dextrose sterilizer
How do we know when a fermenter is complete? And what is the value of this data point?
DX concentration needs to be at 0 g/L, 0.5g/L is acceptable but the lower the better
Draw the hot water system. List all of the systems that utilize this hot water system.
List: broth kill, waste kill, DX heater, sulfate solution heater, mag sulfate heater, citric acid heater
closed loop of RO water that fills this standpipe and heated up with shell and tube HX
Draw the media sterilizer and be able to explain all of the components.
Group 1, 2, RO water, vent, recycle to balance tank, feed pump, reclaim, booster, komax, HTST, reclaim, cooler, ammonia addition to ferms
Draw a seed fermenter and list all of the major components. Be able to explain the purpose of each component.
DX, media, antifoam, CIP, scrubber, offgas, inoc header, chilled water jacket, ammonia, air, steam, sample port, transfer header to prod, DO, pH (2), TT (2), PT (2), LT, Antifoam probes (2)
Identify at least 4 key differences between seed fermenter and production fermenter. BONUS points for any additional differences (100 points for additional differences).
1. physical size/capacity
2. different scrubbers
3. DX/media inlet (top on seeds, side on prod)
4. caustic for pH adjustment on prod
5. seed transfers via air, production transfers via pump
6. 3 seeds, 4 production tanks
Define what OUR stands for and what does it control? How would the OUR change if the fermenter was contaminated?
Oxygen uptake rate - DX control
contaminated - increase OUR
Draw the waste kill HTST and be able to label and explain all components.
1. process waste kill tank
2. scrubber
3. transfer pump, reclaimer, booster pump, heater, HTST, reclaimer, cooler
4. process waste water tank
Draw the dextrose sterilizer and be able to explain all components.
60DS DX tank, pump, DX balance tank, feed pump, reclaim, booster, heater, HTST, reclaim, cooler, sterile day tank, fermentation vessels
*recycle before day tank to balance tank or 60DS tank, recycle after day tank back to balance tank
*cooler - cooling tower water
*heater - hot water set: pump, shell and tube with steam, expansion tank, RO water in
Draw a production fermenter and list all of the major components. Be able to explain the purpose of each component.
DX, media, antifoam, CIP, scrubber, offgas, caustic, chilled water jacket, ammonia, air, steam, sample port, transfer header to surge tanks, DO, pH (2), TT (2), PT (2), LT, Antifoam probes (2)
broth header to pump to surge tanks with blowers to feed pumps that recycle or go to reclaim, heater, HTST, reclaim, and then to ultrafiltration or back to surge tanks
Which process variables can be analyzed continuously in the fermenter vs which have to be analyzed in the lab?
in process: DO, pH, temp, level, backpressures, OUR, CER, foam probes
lab: HPLC, ICP, YSI for DX, conductivity, OD, reference pH